Simultaneous determination of 13 paralytic shellfish toxins and tetrodotoxin in marine shellfish by porous graphitic carbon solid-phase extraction and hydrophilic interaction chromatography-tandem mass spectrometry.

The lethal neurotoxins, paralytic shellfish toxins (PSTs), and tetrodotoxin (TTX) have recently been found in marine shellfish from many coastal states. Herein, we applied a sensitive and reliable ultra-performance hydrophilic interaction chromatography (HILIC)-tandem mass spectrometry (MS) method to determine 13 PSTs and TTX in marine shellfish using a porous carbon solid-phase extraction (SPE). This in-house validation study required the development of a novel chromatographic separation using a HILIC-Z column, which was necessary to retain highly polar compounds. Using acetonitrile as the organic phase and ammonium formate-formic acid buffer as the aqueous phase, the quantitative analysis was carried out with an external standard method in the multiple reaction monitoring modes using positive electrospray ionization. To reduce interference, 1% aqueous acetic acid extracts of the shellfish samples were cleaned up by ion-pair SPE using a porous graphitic carbon cartridge. The calibration curves for PSTs and TTX were linear (R2 > 0.995), and the sensitivity was good, with limits of detection (LODs) of 1.7-13.7 µg/kg, and limits of quantitation (LOQs) of 5.2-41.0 µg/kg. The recoveries were 76.5-95.5% with RSDs of 3.1-12.0%. Finally, We applied the method for the determination of PSTs and TTX in three batches of Nassarius showing excellent method accuracy against expected values.

Immunotoxicity of pentachlorophenol to a marine bivalve species and potential toxification mechanisms underpinning.

The ubiquitous presence of pentachlorophenol (PCP) in ocean environments threatens marine organisms. However, its effects on immunity of marine invertebrates at environmentally realistic levels are still largely unknown. In this study, the immunotoxicity of PCP to a representative bivalve species was evaluated. In addition, its impacts on metabolism, energy supply, detoxification, and oxidative stress status were also analysed by physiological examination as well as comparative transcriptomic and metabolomic analyses to reveal potential mechanisms underpinning. Results illustrated that the immunity of blood clams was evidently hampered upon PCP exposure. Additionally, significant alterations in energy metabolism were detected in PCP-exposed clams. Meanwhile, the expressions of key detoxification genes and the in vivo contents (or activity) of key detoxification enzymes were markedly altered. Exposure to PCP also triggered significant elevations in intracellular ROS and MDA whereas evident suppression of haemocyte viability. The abovementioned findings were further supported by transcriptomic and metabolomic analyses. Our results suggest that PCP may hamper the immunity of the blood clam by (i) constraining the cellular energy supply through disrupting metabolism; and (ii) damaging haemocytes through inducing oxidative stress. Considering the high similarity of immunity among species, many marine invertebrates may be threatened by PCP, which deserves more attention.

Pollution level and health risk assessment of the total petroleum hydrocarbon in marine environment and aquatic products: a case of China.

To evaluate the pollution level and health risk of total petroleum hydrocarbon (TPH), seawater, sediments, and aquatic organisms were sampled from the southern sea area of Zhejiang Province (Yangtze River Delta, China) between 2017 and 2019. TPH was widely present in the aquatic environment and products, and its concentration was highly variable. The average value of pollution index (PI) exceeded 1 from 2017 to 2018, and 45.46-69.19% of seawater samples and 56.87-50.00% of sediment samples were polluted. The results showed significant differences in the TPH concentration in various species of aquatic organisms. The average TPH value in aquatic organisms could be ranked in the order as follows: bivalve > shrimp > crab > fish, further reflecting that the ability to accumulate and metabolize TPH existed differently among aquatic organisms within the same pond aquaculture environment. It was relatively safe to eat aquatic products since the exposure risk index was found to be far below the threshold value in this study. Therefore, it would be prudent to undertake regular monitoring of TPH to ensure effective ecosystem functioning and seafood safety in the southern Zhejiang ocean.

Spirulina phycocyanin induces differential protein expression and apoptosis in SKOV-3 cells.

The present study was designed to determine the effects of phycocyanin (PC) on Human ovarian cancer SKOV-3 cells and the underlying molecular mechanisms of action. The inhibitory effects of PC on the cell proliferation were detected by MTT assay. The IC50 values of PC were 182.0µM and 133.6µM for 24h and 48h exposure, respectively. PC induced apoptosis in SKOV-3 cells was observed by electron microscopy and flow cytometry. The apoptosis rate was increased from 1.6% to 19.8% after PC exposure. The fluorescence intensity of ROS and the activities of Caspase-3, Caspase-8, and Caspase-9 were increased. Differentiated expression protein spots were selected and identified using proteomic techniques. There were 698±73 and 683±79 protein spots resolved in untreated and PC-treated cells, respectively. Forty five differential protein spots were analyzed by MALDI-TOF-MS, including mtSSB, PSME3, and nucleolin. The mRNA expression profiles determined by RT-PCR were consistent with that of the two-dimensional electrophoresis. The decreased proteins such as HSP60, nucleolin, PPase, peroxiredoxin-4 and the increased protein (mtSSB) were identified in SKOV-3 cells after PC treatment, indicating that the effects of PC on tumor cell apoptosis may be relate to multiple target proteins. And the mitochondrial pathway may be the main pathway for PC-induced apoptosis.